Search

The aim of the present study was to elucidate the molecular mechanism of Aiolos in the regulation of B‑cell leukaemia. A lentiviral system was used for overexpression of the Aiolos gene in Nalm‑6 cells to determine the effects of Aiolos on proliferation, apoptosis and the cell cycle. The expression and activation of phosphatase and tensin homolog deleted on chromosome ten (PTEN) and Akt were also investigated. Upregulation of Aiolos inhibited cell growth and arrested an increased number of Nalm‑6 cells at the G0/G1 phase. The apoptotic cell quantities were also significantly lower in the Aiolos‑transfected Nalm‑6 cells. In addition, Aiolos overexpression downregulated PTEN, but increased the expression and phosphorylation of Akt in the Nalm‑6 cells. The Akt inhibitor, Akti‑1/2, reduced the percentage of viable Aiolos‑overexpressed Nalm‑6 cells, however, it had no effect on cell cycle arrest or proliferation. Aiolos upregulation in the Nalm‑6 cells inhibited cell proliferation, suppressed apoptosis and arrested the cell cycle at the G0/G1 phase. Aiolos improved the survival of Nalm‑6 cells via PTEN‑ and Akt‑dependent processes.

Sodium butyrate (NaBu) is a short-chain fatty acid (SCFA), which has been proposed as a potential anticancer agent. Apoptosis-associated speck-like protein (ASC) is a pro-apoptotic signaling factor that is subjected to epigenetic silencing in human cancers. Modulation by the aberrant methylation of CpG islands of ASC is a well-characterized epigenetic mechanism, and the methylation-induced silencing of ASC has been observed in several types of tumors. NaBu induces cell cycle arrest, markers of cell differentiation and apoptosis in colon cancer. NaBu promotes transcriptional activation by relaxing the DNA conformation and displays anti-proliferative and differentiating activity in a wide variety of cancers. Thus, we used NaBu to investigate the relationship between the status of cell proliferation and the re-expression of ASC in colon carcinoma LS174T cells. Our experiments determined ASC re-expression at the protein level using western blotting. In addition, we used reverse transcription-polymerase chain reaction to detect the expression levels of ASC mRNA and an MTT assay to detect the inhibitory rate of cell growth. The apoptosis rate was also detected for further validation of the re-expression of ASC. The results showed that ASC re-expression was significantly increased in the LS174 cells following NaBu treatment in a time- and dose-dependent manner. The expression of ASC also induced the apoptosis of LS174T cells. These results suggest that NaBu plays a role in the reactivation of ASC expression and that the latter promotes the apoptosis of LS174T cells.

Chronic active Epstein-Barr virus infection (CAEBV) is a progressive and life-threatening disease characterized by persistent or recurrent EBV activation. It has been reported that, γδ T cells, a type of cytotoxic lymphocyte, play a critical role in restricting EBV. However, the functional status of γδ T cells in pediatric CAEBV patients has not yet been described. In this study, flow cytometry analysis was conducted to explore the cytokine production capacity of γδ T cells in CAEBV patients. A diminished frequency of γδ T cells and decreased expression of cytolytic molecule granzyme B were found in CAEBV patients, suggesting a dysfunction in the immune regulatory function of γδ T cells in this disease. [ABSTRACT FROM AUTHOR]

Fibrosis is a prevalent pathological condition observed in various organs and tissues. It primarily arises from the excessive and abnormal accumulation of the extracellular matrix, resulting in the structural and functional impairment of tissues and organs, which can culminate in death. Many forms of fibrosis, including liver, cardiac, pulmonary, and renal fibrosis, are considered irreversible. Maternally expressed gene 3 (MEG3) is an imprinted RNA gene. Historically, the downregulation of MEG3 has been linked to tumor pathogenesis. However, recent studies indicate an emerging association of MEG3 with fibrotic diseases. In this review, we delve into the current understanding of MEG3's role in fibrosis, aiming to shed light on the molecular mechanisms of fibrosis and the potential of MEG3 as a novel therapeutic target. [ABSTRACT FROM AUTHOR]

Background: Dermatophytosis is an intractable superficial mycosis in humans and animals mainly caused by Trichophyton mentagrophytes (T. mentagrophytes), with a global prevalence of about 20%. Keratinocytes are the most abundant participants in skin immunity, and they also play a role in the first‐line defence against T. mentagrophytes. However, no studies of keratinocyte responses against T. mentagrophytes infection based on the whole transcriptome have been reported. Objectives: Here, we systematically analysed changes in keratinocytes infected with T. mentagrophytes using whole transcriptome sequencing technology. Methods: The phenotypic changes in keratinocytes after infection with 1 × 105 conidia/mL T. mentagrophytes were observed by light microscopy, scanning electron microscopy, transmission electron microscopy and terminal deoxynucleotidyl transferase dUTP nick end labeling. RNA‐sequencing (RNA‐seq), small RNA‐seq technology and related bioinformatics methods were used to systematically analyse the whole transcriptome changes in keratinocytes upon T. mentagrophytes stimulation. Results: We found that T. mentagrophytes infection caused morphological changes, membrane damage, the formation of irregular organelles and keratinocyte apoptosis. A total of 204 differentially expressed (DE) circular RNAs (circRNAs), 868 DE long noncoding RNAs (lncRNAs), 2973 DE mRNAs and 209 DE micro RNAs (miRNAs) were identified between noninfected and T. mentagrophytes‐infected keratinocytes. The expression level of selected RNAs was validated by quantitative real‐time polymerase chain reaction (qRT‐PCR). Functional enrichment analysis revealed that the parental genes of DE circRNAs were related to cell response, cell death and establishment of the skin barrier. Genes targeted by miRNA were involved in regulating the initiation of the immune response. Based on the expression level of circRNAs, lncRNAs, mRNAs and miRNAs, circRNA‐miRNA‐mRNA competing endogenous (ceRNA) networks comprised of 159 DE miRNAs, 141 DE circRNAs and 2307 DE mRNAs, and lncRNA‐miRNA‐mRNA ceRNA networks comprised of 790 DE lncRNAs, 190 DE miRNAs and 2663 DE mRNAs were constructed. The reliability of two selected ceRNA networks was verified using qRT‐PCR. Further functional enrichment analysis revealed that the DE mRNAs interacting with circRNAs and lncRNAs in the ceRNA network mainly participated in fungal recognition, inflammation, the innate immune response and the death of keratinocytes. Conclusions: Our findings might provide new evidence on the pathogenesis of T. mentagrophytes‐induced dermatophytosis, which is essential for identifying new therapeutic targets for dermatophytosis treatment. [ABSTRACT FROM AUTHOR]

Objective: To explore the impact of pyroptosis related gene GSDME on the prognosis of patients with nasopharyngeal carcinoma and its interaction with the immune microenvironment. Methods: Gene expression profile and clinical data of 548 patients with nasopharyngeal carcinoma were obtained from TCGA database. Differential expression analysis and GO-KEGG enrichment analysis of GSDME were conducted using R language. Immunohistochemical maps of GSDME protein in nasopharyngeal carcinoma tissues and corresponding para-carcinoma tissues were obtained from The Human Protein Atlas database. The interacting proteins of GSDME were explored by using STRING database. The correlation between GSDME expression and infiltration of 24 kinds of immune cells in nasopharyngeal carcinoma tissues was analyzed by ssGSEA algorithm, the correlation between GSDME expression and immune checkpoint molecules was analyzed by Spearman method, and the correlation between GSDME expression and cytokine expression was analyzed by using TISIDB database. Univariate and multivariate COX regression analyses were used to screen the risk factors for prognosis, based on which, the nomogram and calibration curve were drawn. Survival analysis and risk analysis were performed in nasopharyngeal carcinoma patients based on GSDME expression levels. The mRNA expression of GSDME and four chemokines in Chinese nasopharyngeal carcinoma tissues was verified by qPCR. Results: Analysis of database data showed that GSDME was highly expressed in nasopharyngeal carcinoma tissues compared with paraneoplastic tissues (P<0.01), GO-KEGG enrichment analysis showed that GSDME was involved in immune response, cell pyroptosis-related signaling pathways, and GSDME expression was correlated with immune cell infiltration, cytokine and immune checkpoint molecule expression. qPCR examination of Chinese nasopharyngeal carcinoma tissues verified the bioinformatics predictions; GSDME expression, N stage and M stage were risk factors for the prognosis of nasopharyngeal carcinoma patients, and the nomogram and calibration curve based on risk factors could better predict the OS of nasopharyngeal carcinoma patients, and patients with high GSDME expression in nasopharyngeal carcinoma tissues had poor prognosis. Conclusion: GSDME is highly expressed in nasopharyngeal carcinoma tissues and is associated with patient's poor prognosis. High GSDME expression is associated with the pro-cancer effect of the immune microenvironment in nasopharyngeal carcinoma, which may be a potential target for immunotherapy. [ABSTRACT FROM AUTHOR]

TP53 is a well-known tumor suppressor gene and one of the most common genetic alterations in human cancers. However, the role of p53 as a prognostic marker of esophageal squamous cell carcinoma (ESCC) is controversial in the association between TP53 alterations and clinical outcomes. To address this issue, we evaluated TP53 mutations, p53 protein expression, clinicopathological parameters, and survivals rates in a large scale of patients with ESCC. Two cohorts were included in this study: TP53 mutations were detected by next-generation sequencing in 316 ESCC patients, and p53 protein expression was tested by immunohistochemistry in 6,028 ESCC patients. Survival analysis was performed using the Kaplan–Meier curve and the Cox proportional hazards model. TP53 mutations were found in ESCC patients from 241 of 316 (76.3%), and the rate of positive expression of p53 protein was 59.1% in 6,028 ESCC patients (including 1819 with high expression of p53 protein), respectively. Most mutations were missense, which has a high expression of p53 protein. Compared with wild-typeTP53, TP53 gene mutations were not significantly associated with survival time (p = 0.083). In multivariate analysis, the p53 protein expression was an independent prognostic factor for ESCC. The high-expression group of p53 protein has poor survival (p < 0.001) compared to low-expression group in patients with ESCC. The high expression of the p53 protein, not the TP53 mutation, is predictive of poor survival in patients with ESCC, and p53 protein expression might have the potential to be a prognosis biomarker and therapy target in ESCC. [ABSTRACT FROM AUTHOR]

Insect TRPV is one subfamily of transient receptor potential (TRP) channels, and it is composed of Inactive (Iav) and Nanchung (Nan), the molecular targets of afidopyropen in several sucking insect pests. In this study, we performed successive selection and synergism tests based on previous work. The resistant afidopyropen strain HD-Afi of Bemisia tabaci reached about 86-fold resistance to afidopyropen, and only part of the resistance resulted from detoxification of cytochrome P450 monooxygenases. Then, we cloned and characterized Nan and Iav from B. tabaci (BtNan and BtIav), and found that they were expressed in all stages of development and tissues of adult, and the expression level of BtNan in strains of HD-S and HD-Afi were significantly increased after afidopyropen treatment. Further, expression of BtNan was downregulated after gene silencing, and it resulted in significantly decreased afidopyropen resistance in the strain of HD-Afi. Our data revealed the first evidence that overexpression of the TRPV Nan gene is responsible for causing afidopyropen resistance in B. tabaci , and our results could provide new visions on the function of TRPVs in the development of resistance to pesticide and supply evidence for developing novel strategies of pest management. [ABSTRACT FROM AUTHOR]

Simple Summary: The fall armyworm (FAW), Spodoptera frugiperda, is a serious worldwide agricultural pest, threatening food security and crop production. Sex pheromone lures are commonly used in population monitoring and biological control of FAWs. Although the sex pheromone components of the FAW have been successfully identified, there are no reports on the molecular mechanism of FAW sex pheromone biosynthesis. In this study, we identified a total of 99 genes related to the biosynthesis of sex pheromones from the S. frugiperda genome, which belonged to 11 families of genes. Based on gene expression patterns and phylogenetic analysis, several genes had PG-biased expression, indicating that they may play an important role in sex pheromone biosynthesis. These results could lay a solid foundation for understanding the molecular mechanisms of S. frugiperda sex pheromone biosynthesis and provide new targets for developing novel pest control methods based on disrupting sexual communication. Spodoptera frugiperda is an agricultural pest causing substantial damage and losses to commercial crops. Sex pheromones are critical for successful mating in Lepidoptera and have been used for monitoring and control of many pest species. The sex pheromone of S. frugiperda is known, but the genes involved in its biosynthesis have not been identified. We systematically studied 99 candidate sex pheromone genes in the genome of S. frugiperda including 1 acetyl-CoA carboxylase (ACC), 11 fatty acid synthases (FASs), 17 desaturases (DESs), 4 fatty acid transport proteins (FATPs), 29 fatty acyl-CoA reductases (FARs), 17 acetyl-CoA acetyltransferases (ACTs), 5 acyl-CoA dehydrogenase (ACDs), 3 enoyl-CoA hydratases (ECHs), 3 hydroxyacyl-CoA dehydrogenases (HCDs), 6 ethyl-CoA thiolases (KCTs), and 3 acyl-CoA-binding proteins (ACBPs). Based on the comparative transcriptome results, we found 22 candidate sex pheromone biosynthesis genes predominately expressed in pheromone glands (PGs) than abdomens without PGs including SfruFAS4, SfruFATP3, SfruACD5, SfruKCT3, SfruDES2, SfruDES5, SfruDES11, SfruDES13, SfruFAR1, SfruFAR2, SfruFAR3, SfruFAR6, SfruFAR7, SfruFAR8, SfruFAR9, SfruFAR10, SfruFAR11, SfruFAR14, SfruFAR16, SfruFAR29, SfruACT6, and SfruACT10. A combination of phylogenetic and tissue-specific transcriptomic analyses indicated that SfruDES5, SfruDES11, SfruFAR2, SfruFAR3, and SfruFAR9 may be key genes involved in the sex pheromone synthesis of S. frugiperda. Our results could provide a theoretical basis for understanding the molecular mechanisms of sex pheromone biosynthesis in S. frugiperda, and also provide new targets for developing novel pest control methods based on disrupting sexual communication. [ABSTRACT FROM AUTHOR]

Glioblastoma (GBM) is the most common primary malignant brain tumor in adults. Temozolomide (TMZ) is used as the standard chemotherapeutic agent for GBM but with limited success, and treatment failure is mainly due to tumor resistance. One of the leading causes of TMZ resistance is the upregulation of the DNA repair mechanism. Therefore, targeting the DNA damage response (DDR) is proposed to be an effective strategy to sensitize tumor cells to TMZ. In the present study, we demonstrated that the combined use of the PI3K inhibitor ZSTK474 and TMZ showed synergetic anticancer effects on human GBM cells in vitro and in vivo. The combination treatment led to significantly increased cell apoptosis and DNA double strand breaks (DSBs). In addition, a mechanistic study indicated that TMZ enhanced the homologous recombination (HR) repair efficiency in GBM cells, while ZSTK474 impaired HR repair by blocking the phosphorylation of ATM and the expression of BRCA1/2 and Rad51, thereby sensitizing GBM cells to TMZ. Moreover, TMZ activated the PI3K signaling pathway through upregulation of the PI3K catalytic subunits p110α and p110β and the phosphorylation of Akt. Meanwhile, ZSTK474 blocked the activity of the PI3K/Akt pathway. Taken together, our findings suggested that the combination of ZSTK474 and TMZ might be a potential therapeutic option for GBM. [ABSTRACT FROM AUTHOR]

Soybean Sclerotinia stem rot is caused by Sclerotinia sclerotiorum infection, which causes extensive and severe damage to soybean production. Here, we isolated and patented a Bacillus amyloliquefaciens strain, and used it to verify the antagonistic effect of B. amyloliquefaciens on S. sclerotiorum and to explore the possible underlying mechanism. First, we conducted a plate confrontation experiment using the two microbes. Then, inoculation of soybean (Glycine max) seedlings with S. sclerotiorum (Gm-Ss), B. amyloliquefaciens (Gm-Ba), and their combination (Gm-Ba-Ss) was performed, followed by dual RNA sequencing analysis. Plate confrontation and inoculation experiments showed that B. amyloliquefaciens significantly antagonized S. sclerotiorum growth. The average number of fragments per kilobase of transcript per million fragments mapped of S. sclerotiorum transcripts in Gm-Ss and Gm-Ba-Ss inoculation treatments were 117.82 and 50.79, respectively, indicating that B. amyloliquefaciens strongly inhibited gene expression of S. sclerotiorum. In contrast, the average number of fragments per kilobase of transcript per million fragments mapped of B. amyloliquefaciens transcripts in Gm-Ba and Gm-Ba-Ss inoculation treatments were 479.56 and 579.66, respectively, indicating that S. sclerotiorum promoted overall gene expression in B. amyloliquefaciens. For S. sclerotiorum , 507 upregulated and 4,950 downregulated genes were identified among 8,975 genes in the paired comparison Gm-Ba-Ss vs. Gm-Ss. These differentially expressed genes (DEGs) were significantly enriched in the ribosome (ko03010) KEGG pathway. Additionally, for B. amyloliquefaciens , 294 upregulated and 178 downregulated genes were identified among all 3,154 genes in the paired comparison Gm-Ba-Ss vs. Gm-Ba, and these DEGs were mainly and significantly enriched in metabolism-related KEGG pathways, including the citrate cycle (ko00020) and carbon metabolism (ko01200). We concluded that B. amyloliquefaciens inhibits the expression of genes encoding the ribosomal subunit of S. sclerotiorum , resulting in protein synthesis inhibition in S. sclerotiorum , and thus had a strong antagonistic effect on the fungus. This study provides a scientific basis for the biological control of S. sclerotiorum by B. amyloliquefaciens. [ABSTRACT FROM AUTHOR]

A growing number of studies have focused on the microRNA (miRNA) expression in Bemisia tabaci , one devastating agricultural insect pest of the tropical and subtropical areas for which the primary means of control are insecticides. In studying the genetic underpinnings of insecticide resistance, the choice of stable reference genes for normalizing data plays a key role to acquire unbiased expression profile results from quantitative real-time PCR (qPCR) analysis. Expression profiles of 11 selected reference genes were determined systematically in B. tabaci exposure to 11 insecticides. Furthermore, we assessed the stability of all the selected candidates in relation to other variables including sex, tissue type, and developmental stage. Candidate reference gene validation was conducted by analyzing the let-7-5p expression under various experimental treatments. Five programs BestKeeper, NormFinder, geNorm, △Ct, and RefFinder were applied to verify the accuracy of the selected candidates. Our results displayed that the best choices of the selected candidates for pymetrozine, sulfoxaflor, flonicamid, cyantraniliprole, afidopyropen, and deltamethrin treatment were miR-1-3p and miR-100-5p, U6 and miR-100-5p were best for chlorpyrifos and imidacloprid treatments, and U6 and miR-1-3p were best for flupyradifurone and β-cypermethrin treatments. The reference genes miR-624, miR-252, and miR-275 worked best in adult tissues, miR-100-5p and miR-1-3p worked best in either sex, and miR-624 and miR-11 were best to use across developmental stages. Not even one reference gene was found to be suitable for all experimental conditions. Our results contributed to the growing body of the literature on qPCR reference gene selection under various experimental conditions and facilitate further investigation on gene expression changes in B. tabaci , resulting from pesticide exposure. [ABSTRACT FROM AUTHOR]

The advances in single-cell RNA sequencing (scRNA-seq) technologies enable the characterization of transcriptomic profiles at the cellular level and demonstrate great promise in bulk sample analysis thereby offering opportunities to transfer gene signature from scRNA-seq to bulk data. However, the gene expression signatures identified from single cells are typically inapplicable to bulk RNA-seq data due to the profiling differences of distinct sequencing technologies. Here, we propose single-cell pair-wise gene expression (scPAGE), a novel method to develop single-cell gene pair signatures (scGPSs) that were beneficial to bulk RNA-seq classification to transfer knowledge across platforms. PAGE was adopted to tackle the challenge of profiling differences. We applied the method to acute myeloid leukemia (AML) and identified the scGPS from mouse scRNA-seq that allowed discriminating between AML and control cells. The scGPS was validated in bulk RNA-seq datasets and demonstrated better performance (average area under the curve [AUC] = 0.96) than the conventional gene expression strategies (average AUC |$\le$|  0.88) suggesting its potential in disclosing the molecular mechanism of AML. The scGPS also outperformed its bulk counterpart, which highlighted the benefit of gene signature transfer. Furthermore, we confirmed the utility of scPAGE in sepsis as an example of other disease scenarios. scPAGE leveraged the advantages of single-cell profiles to enhance the analysis of bulk samples revealing great potential of transferring knowledge from single-cell to bulk transcriptome studies. [ABSTRACT FROM AUTHOR]

As important regulators, miRNAs could play pivotal roles in regulation of fruit coloring. Actinidia arguta is a newly emerged fruit tree with extensively application prospects. However, miRNAs involved in A. arguta fruit coloring are unknown. In this study, A. arguta fruit were investigated at three developmental stages by small RNAs high-throughput sequencing. A total of 482 conserved miRNAs corresponding to 526 pre-miRNAs and 581 novel miRNAs corresponding to 619 pre-miRNAs were grouped into 46 miRNA families. Target gene prediction and analysis revealed that miR858, a strongly candidate miRNA, was involved in anthocyanin biosynthesis in which contributes to fruit coloring. The anthocyanin level was determined in three A. arguta cultivars by UPLC-MS/MS (ultra-performance liquid chromatography coupled with tandem mass spectrometry). In addition, qPCR (quantitative real-time PCR), cluster analysis were conducted as well as correlation analysis. All results were combined to propose a model in which describes an association of miRNA and anthocyanin biosynthesis in A. arguta. The data presented herein is the first report on miRNA profile analysis in A. arguta, which can provide valuable information for further research into the regulation of the miRNAs in anthocyanin biosynthesis and fruit coloring. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are small noncoding RNAs. These noncoding RNAs regulate the expression of target genes and inhibit the translation of target proteins at the post-transcriptional level. miRNAs also play an important role in human health, from the development and differentiation of cells to the occurrence and progression of disorders such as cancer, cardiovascular diseases, and neurodegenerative diseases. Precancerous lesions are lesions prior to invasive carcinomas, and carcinogenesis is a very complicated process, which is multistage and the result of multigene synergy. miRNAs exert effects as both oncogenes and tumor suppressor genes by regulating target genes involved in signaling pathways. Hence, precancerous lesions are accompanied by relevant miRNA changes. Based on the morphology of miRNAs in vivo and the specificity of miRNA, various novel miRNA analysis methods have been developed, including reverse transcription quantitative PCR, enzyme analysis, molecular beacons, and deep sequencing. For example, in the laryngeal epithelial precancerous lesions, the data demonstrate that the expression of miR-10a-5p is downregulated and miR-484 is the most abundant miRNA in hepatic precancerous lesions. In this review, we discuss the functional roles of miRNAs in human precancerous lesions. [ABSTRACT FROM AUTHOR]

Harmonia axyridis (Coleoptera: Coccinellidae) is a polyphagous insect that is an important biological agent used to control agricultural and forestry pests. The role of functional genes in H. axyridis based on quantitative real-time PCR (qRT-PCR) is increasingly well understood to investigate biology, physiology, feeding behavior and the role of important genes in physiological processes. Quantitative real-time PCR (qRT-PCR) is a powerful and reliable technique to quantify gene expression. Using qRT-PCR, expression levels of target genes are determined based on the levels of internal reference genes; therefore, reference genes need to be stably expressed under specific experimental conditions. However, there have been no studies on the stability of reference genes used in H. axyridis. In this study, we systematically investigated expression profiles of nine candidate reference genes from H. axyridis, including β-actin (ACTIN); elongation factor 1 α (EF1A); ribosomal proteins L10, L18, L28, S13, and S15 (RPL10, RPL18, RPL28, RPS13 and RPS15); glyceraldehyde-3-phosphate dehydrogenase (GAPDH); and superoxide dismutase (SOD). Four analytical methods (geNorm, BestKeeper, NormFinder, and the ΔCt method) were used to evaluate the suitability of these genes as internal reference genes for three biotic factors (developmental stage, tissue, and sex) and two abiotic treatments (temperature and photoperiod). RefFinder, a comprehensive evaluation platform integrating the four analytical methods, was used to rank the overall stability of these reference genes. Among the nine candidate genes, different reference genes were identified as having the most stable expression across biotic and abiotic factors. Genes encoding ribosomal proteins typically had the most stable expression, though EF1A was the most stable across developmental stages and photoperiods. To validate the suitability of these reference genes, heat shock protein 90 (HSP90) was chosen as a target. Significant up-regulation in HSP90 expression level in response to both low and high temperature was observed when using the most suitable reference genes but not when using an arbitrarily selected reference gene. The reference genes identified in this study will provide the basis for future functional genomics research in H. axyridis and will also facilitate the establishment of a standardized qRT-PCR program for other related insects. [ABSTRACT FROM AUTHOR]

B-box containing proteins (BBXs) are a class of zinc finger transcription factor proteins that contain one or two B-box domains and play important roles in plant growth and development. The BBX gene family has been identified in theArabidopsisand rice genomes. However, no comprehensive identification has been reported in pear. In this study, 37 genes were identified as candidateBBXs in Chinese white pear (Pyrus bretschneideriRehd) and denominated following the uniform nomenclature inArabidopsis. We also performed analyses of their phylogenic relationships, structures, chromosomal locations, promoter regions, Gene Ontology (GO) annotations, and expression patterns. The identified genes were classified into five groups based on phylogeny and domain constitution analysis.PbBBXswere unevenly distributed in 12 out of 17 pear chromosomes. Thecis-elements in the promoters ofPbBBXs were mainly related to light, stress, and hormone response. Gene expression analysis by quantitative real-time PCR showed thatPbBBXs had differential expression in different plant organs and could be classified into four major groups.PbBBXs also showed differential responses to low temperature and dehydration treatments. Overall, this study provided useful information for further functional analysis ofBBXs in pear. [ABSTRACT FROM PUBLISHER]

Imidacloprid and sulfoxaflor have potential damage to nontarget aquatic organisms. However, limited information has been provided on their underlying toxicity effects on Rana nigromaculata tadpoles. Thus, the acute toxicity and chronic effects of imidacloprid and sulfoxaflor on R. nigromaculata tadpoles were studied. Acute toxicity indicted that 96 h for LC 50 values of imidacloprid and sulfoxaflor were 173.55 and 427.37 mg/L, respectively. In this research, we explored antioxidant enzymes, some biological indexes, hormone levels and expression of relative tadpole genes involved in thyroid hormone-dependent metaplastic development after exposure for 28 days under 1/10 and 1/100 LC 50. Results showed an increase in the activity of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) in the tadpoles, and a low bioconcentration level with a bioconcentration factor (BCFs) < 1. The impact on the development of tadpoles was induced by the breakdown of hormonal levels engaged in metamorphosis. According to the real-time PCR results, imidacloprid and sulfoxaflor delayed amphibian metamorphosis by modifying mRNA expression, indicating that imidacloprid and sulfoxaflor may have an endocrine-disrupting effect on R. nigromaculata tadpoles. These findings were indicative of the toxicity of imidacloprid and sulfoxaflor to R. nigromaculata tadpoles. [ABSTRACT FROM AUTHOR]

The sycamore lace bug, Corythucha ciliata (Hemiptera: Tingidae), is an invasive forestry pest rapidly expanding in many countries. This pest poses a considerable threat to the urban forestry ecosystem, especially to Platanus spp. However, its molecular biology and biochemistry are poorly understood. This study reports the first C. ciliata transcriptome, encompassing three different life stages (Nymphs, adults female (AF) and adults male (AM)). In total, 26.53 GB of clean data and 60,879 unigenes were obtained from three RNA-seq libraries. These unigenes were annotated and classified by Nr (NCBI non-redundant protein sequences), Nt (NCBI non-redundant nucleotide sequences), Pfam (Protein family), KOG/COG (Clusters of Orthologous Groups of proteins), Swiss-Prot (A manually annotated and reviewed protein sequence database), and KO (KEGG Ortholog database). After all pairwise comparisons between these three different samples, a large number of differentially expressed genes were revealed. The dramatic differences in global gene expression profiles were found between distinct life stages (nymphs and AF, nymphs and AM) and sex difference (AF and AM), with some of the significantly differentially expressed genes (DEGs) being related to metamorphosis, digestion, immune and sex difference. The different express of unigenes were validated through quantitative Real-Time PCR (qRT-PCR) for 16 randomly selected unigenes. In addition, 17,462 potential simple sequence repeat molecular markers were identified in these transcriptome resources. These comprehensive C. ciliata transcriptomic information can be utilized to promote the development of environmentally friendly methodologies to disrupt the processes of metamorphosis, digestion, immune and sex differences. [ABSTRACT FROM AUTHOR]

The whitefly Bemisia tabaci is a genetically diverse complex with multiple cryptic species, and some are the most destructive invasive pests of many ornamentals and crops worldwide. Encarsia sophia is an autoparasitoid wasp that demonstrated high efficiency as bio-control agent of whiteflies. However, the immune mechanism of B. tabaci parasitization by E. sophia is unknown. In order to investigate immune response of B. tabaci to E. Sophia parasitization, the transcriptome of E. sophia parasitized B. tabaci nymph was sequenced by Illumina sequencing. De novo assembly generated 393,063 unigenes with average length of 616 bp, in which 46,406 unigenes (15.8% of all unigenes) were successfully mapped. Parasitization by E. sophia had significant effects on the transcriptome profile of B. tabaci nymph. A total of 1482 genes were significantly differentially expressed, of which 852 genes were up-regulated and 630 genes were down-regulated. These genes were mainly involved in immune response, development, metabolism and host signaling pathways. At least 52 genes were found to be involved in the host immune response, 33 genes were involved in the development process, and 29 genes were involved in host metabolism. Taken together, the assembled and annotated transcriptome sequences provided a valuable genomic resource for further understanding the molecular mechanism of immune response of B. tabaci parasitization by E. sophia. [ABSTRACT FROM AUTHOR]

We isolated JEUD-38, a new sesquiterpene lactone from Inula japonica Thunb. JEUD-38 dramatically attenuated lipopolysaccharide (LPS)-induced nitric oxide (NO) production. Consistent with this finding, the protein expression of inducible nitric oxide synthase (iNOS) was blocked by JEUD-38 in a concentration-dependent manner. To elucidate the mechanism, we examined the effect of JEUD-38 on LPS-stimulated nuclear factor-κB (NF-κB) nuclear translocation, inhibitory factor-κB (IκB) phosphorylation, and degradation. JEUD-38 reduced the translocation of p65, via abrogating IκB-α phosphorylation and degradation. In addition, JEUD-38 inhibited LPS-stimulated phosphorylation of mitogen-activated protein kinases (MAPKs) including extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38. Since iNOS as well as the upstream NF-κB and MAPKs are known to be closely involved in inflammation, these results suggest that JEUD-38 is a promising candidate for prevention and therapy of inflammatory diseases. [ABSTRACT FROM AUTHOR]

Background/Aims: Golgi phosphoprotein 3 (GOLPH3) is a newly reported oncogene that plays a significant role in regulating cell growth. Recent research has shown that overexpression of GOLPH3 is correlated with patient survival and M classification in breast cancer and other cancers. However, the mechanisms by which GOLPH3 contributes to metastasis in non-small cell lung cancer (NSCLC) have not been previously clarified and are therefore the focus of this work. Methods: Immunohistochemistry (IHC) and western blotting analysis were performed to assess the GOLPH3 protein level, small interfering RNA (siRNA) and transwell assays were conducted to investigate the role of GOLPH3 in migration and invasion, and real-time PCR was performed to estimate the level of GOLPH3 mRNA expression. Results: GOLPH3 was significantly correlated with clinicopathological variables, such as the clinical stage (P=0.012), T classification (P=0.002) and metastasis (M classification) (P=0.008), in NSCLC patients and was negatively correlated with the prognosis. Knockdown of GOLPH3 significantly suppressed the migratory and invasive ability of NSCLC cell lines and downregulated the enzyme activity and protein levels of MMP-2 and MMP-9. Conclusions: The expression level of GOLPH3 is correlated with metastasis and prognosis in NSCLC, and GOLPH3 mediates metastasis by regulating the protein levels of MMP-2 and MMP-9 in vitro. © 2015 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Introduction The upregulation of Nav1.8 in primary afferents plays a critical role in the development and persistence of neuropathic pain. The mechanisms underlying the upregulation are not fully understood. Aims The present study aims to investigate the regulatory effect of histamine on the expression of Nav1.8 in primary afferent neurons and its involvement in neuropathic pain. Results Histamine at 10−8 M increased the expression of Nav1.8 in cultured DRG neurons. This effect could be blocked by H2 receptor antagonist cimetidine or famotidine, but not by H1 receptor antagonist pyrilamine or dual H3/H4 antagonist thioperamide. Peri-sciatic administration of histamine increased Nav1.8 expression in the sciatic nerve and L4/L5 DRG neurons in a dose-dependent manner, accompanied with remarkable mechanical allodynia and heat hyperalgesia in the ipsilateral hindpaw. Famotidine but not pyrilamine or thioperamide inhibited Nav1.8 upregulation and pain hypersensitivity. In addition, famotidine (40 mg/kg, i.p.) not only suppressed autotomy behavior in the rat neuroma model of neuropathic pain but also attenuated mechanical allodynia and thermal hyperalgesia following partial sciatic nerve ligation. Moreover, famotidine inhibited Nav1.8 upregulation in the neuroma and ligated sciatic nerve. Conclusions Our findings indicate that histamine increases Nav1.8 expression in primary afferent neurons via H2 receptor-mediated pathway and thereby contributes to neuropathic pain. H2 receptor antagonists may potentially be used as analgesics for patients with neuropathic pain. [ABSTRACT FROM AUTHOR]

Abstract: Notch signaling is often involved in early development which helps to determine the differentiation state and fate of stem cells destined to form different tissues in the body. Its role in the differentiation of BM-MSCs (bone marrow-derived mesenchymal stem cells) is much less clear. As there is great interest in the potential of BM-MSCs as a source of cells for treating liver damage, it is important to understand if Notch signaling promotes or suppresses BM-MSCs differentiation into hepatocytes. In the present study, RT-PCR, Western blot results and morphologic changes demonstrated that BM-MSCs could successfully differentiate into hepatocytes in our special induction system including the tissue extract of damaged liver. On the 21st day when the differentiation direction was determined in BM-MSCs, the mRNA level of Jagged2, Delta1, Delta3, Notch1, Notch2, Notch3 and Presenilin1, was significantly lower than that on days 0, 7, and 11. In the further experiments, down-regulation of Notch signaling was shown to be critical for BM-MSCs to differentiate into hepatocytes, as increased Jagged1 resulted in up-regulated Notch activation leading to higher levels of expression of Hes1 and Hey1, which completely blocked Albumin expresion in BM-MSCs. These results in our study showed that Notch signaling in BM-MSCs was necessary to initiate differentiation into hepatocytes, but must be down-regulated for the differentiation to proceed continuously. [Copyright &y& Elsevier]

Objective: To investigate the synergistic effects of carnosic acid (CA) with arsenic trioxide (AsO) on proliferation and apoptosis in HL-60 human myeloid leukemia cells, and the major cellular signaling pathway involved in these effects. Methods: HL-60 cellular proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. Cell cycle distribution and apoptosis were monitored by flow cytometry. The activation of casepase-9, Bcl-2-associated agonist of cell death (BAD), p-BAD, p27, phosphatase and tensin homolog deleted on chromosome ten (PTEN), Akt, p-Akt was assessed by Western blot analysis. The expression of PTEN mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR) analysis. Results: CA reduced HL-60 cell viability in a dose- and time-dependent manner, and induced G1 arrest and apoptosis. Moreover, CA upregulated PTEN expression, blocked the Akt signaling pathway, subsequently inhibited phosphorylation of BAD, reactivated caspase-9, and elevated levels of p27. CA also augmented these effects of AsO. Conclusion: CA might be a novel candidate of the combination therapy for leukemia treatment; these effects were apparently associated with the modulation of PTEN/Akt signaling pathway. [ABSTRACT FROM AUTHOR]

Abstract: Cdc20 is a co-activator of the anaphase-promoting complex/cyclosome (APC/C complex), which recruits substrates at particular phases of the cell cycle and mediates their degradation. Sp100 is a PML-NB scaffold protein, which localizes to nuclear particles during interphase and disperses from them during mitosis, participates in viral resistance, transcriptional regulation, and apoptosis. However, its metabolism during the cell cycle has not yet been fully characterized. We found a putative D-box in Sp100 using the Eukaryotic Linear Motif (ELM) predictor database. The putative D-box of Sp100 was verified by mutational analysis. Overexpression of Cdc20 resulted in decreased levels of both endogenous Sp100 protein and overexpressed Sp100 mRNA in HEK 293 cells. Only an overexpressed D-box deletion mutant of Sp100 accumulated in HEK293 cells that also overexpressed Cdc20. Cdc20 knockdown by cdc20 specific siRNA resulted in increased Sp100 protein levels in cells. Furthermore, we discovered that the Cdc20 mediated degradation of Sp100 is diminished by the proteasome inhibitor MG132, which suggests that the ubiquitination pathway is involved in this process. However, unlike the other Cdc20 substrates, which display oscillating protein levels, the level of Sp100 protein remains constant throughout the cell cycle. Additionally, both overexpression and knockdown of endogenous Sp100 had no effect on the cell cycle. Our results suggested that sp100 is a novel substrate of Cdc20 and it is degraded by the ubiquitination pathway. The intact D-box of Sp100 was necessary for this process. These findings expand our knowledge of both Sp100 and Cdc20 as well as their role in ubiquitination. [Copyright &y& Elsevier]

Red pear is a popular fruit that is appreciated for its attractive and distinctive appearance and mild flavor. In this study, we investigated the mechanism underlying the red coloration of pear skin using the 'Xinqihong' cultivar—which was selected as a spontaneous bud sport mutant of the 'Xinli 7′(Pyrus betulifolia Rehd.) variety and has a stronger red color that is retained in the mature fruit—as an experimental model. We compared the phenotype and gene expression patterns of the two varieties and found no significant differences at the early stage of fruit development. However, although the red color of 'Xinli 7′ fruits began to fade 107 days after full bloom, that of 'Xinqihong' fruits persisted until the time of harvest. Transcriptome sequencing identified 639 genes that were differentially expressed between the two varieties, including genes related to light, calcium, and hormone signaling (e.g., PbPIF3, PbGA2ox, and the calmodulin related genes). Moreover, anthocyanin biosynthesis genes were downregulated as the red color of 'Xinli 7′ fruits faded. These results provide insight into the molecular basis of color differences in red pears that can be useful for improving its fruit quality and commercial value. [ABSTRACT FROM AUTHOR]

Low temperature causes injuries to plants during winter, thereby it affects kiwi fruit quality and yield. However, the changes in metabolites and gene expression during cold acclimation (CA) and deacclimation (DA) in kiwi fruit remain largely unknown. In this study, freezing tolerance, carbohydrate metabolism, and β-amylase gene expression in two Actinidia arguta cv. "CJ-1" and "RB-3" were detected from CA to DA stages. In all acclimation stages, the "CJ-1" was hardier than "RB-3" and possessed lower semi-lethal temperature (LT50). Furthermore, "CJ-1" had a more rapid acclimation speed than "RB-3". Changes of starch, β-amylase, and soluble sugars were associated with freezing tolerance in both cultivars. Starch contents continued to follow a declining trend, while soluble sugars contents continuously accumulated in both cultivars during CA stages (from October to January). To investigate the possible molecular mechanism underlying cold response in A. arguta, in total, 16 AcBAMs genes for β-amylase were identified in the kiwi fruit genome. We carried out localization of chromosome, gene structure, the conserved motif, and the analysis of events in the duplication of genes from AcBAMs. Finally, a strong candidate gene named AaBAM3 from AcBAMs was cloned in Actinidia arguta (A. arguta), The real-time qPCR showed that AaBAM3 gene expression in seasonal changes was consistent with changes of soluble sugars. These results reveal that AaBAM3 may enhance the freezing tolerance of A. arguta through increasing soluble sugar content. [ABSTRACT FROM AUTHOR]

Two distinct clones having high nucleotide identity to the sequences encoding ascorbate peroxidase (APX) were isolated from broccoli (Brassica oleracea L. var. italica). Deduced amino acid sequences of both cDNAs, BO-APX 1 (accession number AB078599) and BO-APX 2 (accession number AB078600), shared identity of 92.8% and there was more than 80% identity between BO-APXs and other plant cytosolic APXs at the protein level. Gene expression and protein levels of BO-APX 1 and BO-APX 2 were investigated in various parts of broccoli after harvest. Transcript levels of BO-APX 2 gradually increased in florets, while those of BO-APX 1 decreased in florets after harvest. BO-APX 1 and BO-APX 2 were expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST) and purified to homogeneity by glutathione sepharose 4B column chromatography. Both proteins of BO-APX 1 and BO-APX 2 appeared as a single major band on SDS-PAGE corresponding to a mass of 25 kDa and reacted with polyclonal antibodies raised against recombinant BO-APX 1. Both enzymes showed high specificities for ascorbate and hydrogen peroxide. The km values of recombinant BO-APX 1 and BO-APX 2 for ascorbate were 395 and 526 μM and those for hydrogen peroxide were 15 and 7 μM, respectively. The role of APX was discussed in relation to ascorbate breakdown in broccoli florets during senescence. [Copyright &y& Elsevier]

The sugarcane shoot borer Chilo infuscatellus (Snellen) is known for causing severe damage to sugarcane yield in China. Methods have been developed to control this pest, including Cry toxin pesticide and transgenic Bt plants. In order to investigate the molecular mechanism of the Cry toxin binding process and provide a basis for understanding the insect's resistance mechanism, we used a high throughput sequencing platform to perform a de novo transcriptome assembly across different larval developmental stages and analyzed Cry toxin receptors based on our assembled transcripts. We cloned twelve Cry toxin receptor genes including 1 cadherin (Cad), 7 aminopeptidase-Ns (APNs), 3 alkaline phosphatases (ALPs), and 1 ATP-binding cassette transporter subfamily C2 (ABCC2), and three of them with full length. The sublethal dosage of Cry1Ac toxin was applied to sugarcane shoot borer and identified some Cry toxin receptor genes that were significantly induced after 48 h of exposure. Furthermore, quantitative RT-PCR was conducted to detect the expression profiles of these genes. Our transcriptome sequence data provided a valuable molecular resource for further study and the identified Cry toxin receptor data gave insights for improved research into the mechanism of Bt resistance. Unlabelled Image • A total of 95,921 unigenes were generated in the de novo Chilo infuscatellus assembled transcriptome. • One hundred and thirty one unigenes were annotated as Cry-toxin receptor homologs. • Twelve Cry-toxin receptor genes were cloned and the expression profiles were also detected. • The sublethal dosage of Cry1Ac toxin induced nine cloned Cry-toxin receptor gene expression. [ABSTRACT FROM AUTHOR]

Abstract The chloride channel (CLC) protein family, which includes both chloride (Cl−) channels and chloride/proton (Cl−/H+) antiporters, is present in all domains of life, from prokaryotes to eukaryotes. However, there are no reported studies about this gene family in tobacco, an economically important global crop plant. In this study, we identified seventeen CLC genes in the genome of Nicotiana tabacum. A multiple sequence alignment showed that all of the predicted proteins shared a high sequence similarity and had a highly conserved GKxGPxxH motif. A gene structure analysis revealed that the NtCLC genes had highly divergent intron-exon patterns. A phylogenetic and conserved motif analysis revealed that the NtCLC family was divided into two clades, in a manner similar to other plants. We also evaluated the expression patterns of these NtCLC genes in different tissues and in plants treated with salt stress. The NtCLC genes had highly variable expression patterns, for example, the largely stem- and bud-specific expression patterns of NtCLC6 and NtCLC8 , respectively. Salt stress treatment (300 mM NaCl) induced the expression of NtCLC2 , NtCLC3 , and NtCLC12 , suggesting that these genes might play a role in tobacco responses to salt stress. Furthermore, the concentration of Cl− in the NtCLC2 - and NtCLC13 -silenced plants showed an obvious lower and higher level, respectively, than the control plants. Thus, we indicated that NtCLC2 or NtCLC13 might play an important role in chloride transport or metabolism in tobacco. Together, these findings establish an empirical foundation for the further functional characterization of the NtCLC genes in tobacco. Highlights • A complete set of 17 members of the CLC family in N. tabacum have been identified in tobacco. • The bioinformatic analyses and expression profiles of NtCLCs in tobacco have been provided. • The expressions of NtCLC2 , NtCLC3 , and NtCLC12 in roots or shoots were induced under salt stress. • NtCLC2 or NtCLC13 might act important role in chloride transport or metabolism in tobacco. [ABSTRACT FROM AUTHOR]

Summary Gastrulation of the mouse embryo entails progressive restriction of lineage potency and the organization of the lineage progenitors into a body plan. Here we performed a high-resolution RNA sequencing analysis on single mid-gastrulation mouse embryos to collate a spatial transcriptome that correlated with the regionalization of cell fates in the embryo. 3D rendition of the quantitative data enabled the visualization of the spatial pattern of all expressing genes in the epiblast in a digital whole-mount in situ format. The dataset also identified genes that (1) are co-expressed in a specific cell population, (2) display similar global pattern of expression, (3) have lineage markers, (4) mark domains of transcriptional and signaling activity associated with cell fates, and (5) can be used as zip codes for mapping the position of single cells isolated from the mid-gastrula stage embryo and the embryo-derived stem cells to the equivalent epiblast cells for delineating their prospective cell fates. [ABSTRACT FROM AUTHOR]

Abstract: Accumulating evidence indicated that smoking might directly induce pulmonary vascular remodeling at the initial stage of chronic obstructive pulmonary disease (COPD). However, the molecular mechanism underlying this process remains poorly understood. To investigate the role of cyclin D1 in pulmonary vascular remodeling, we constructed a plasmid-based short hairpin RNA (shRNA) to knock down the expression of cyclin D1 in smoking rats. Specific shRNA against cyclin D1 significantly prevented the cyclin D1 expression and the cell proliferation in rat pulmonary artery smooth muscle cells (rPASMCs). Furthermore, the plasmid-based shRNA successfully decreased the cyclin D1 protein in intra-pulmonary arteries of smoking rats and subsequently decreased the wall thickness of pulmonary vessels and the percentage of muscularized vessels. We conclude that the plasmid-based shRNA against cyclin D1 gene attenuated pulmonary vascular remodeling in smoking rats. Cyclin D1 might play a critical role in cigarette smoke-induced pulmonary vascular remodeling via regulating rPASMCs proliferation. [Copyright &y& Elsevier]

• Colchicine increased DNA methylation level. • DNA methylation level of pear polyploids is higher than diploids. • DNA methylation may cause changes in plant polyploid phenotype. Polyploid plants of fruit trees are generally characterized by dwarf growth, larger organs, and strong resistance to disease. They are therefore of great value in fruit breeding. Plant epigenetics generally has the forms of DNA methylation, RNA interference, and histone modification. The mechanisms of phenotypic variation and the physiological characteristics of plant polyploids are closely related to epigenetics. However, there have been few studies examining the relationship between DNA methylation and polyploidization in perennial woody plants. In this study, we compared the DNA methylation of unconverted diploids (q2x-1), tetraploids (4x-1, 4x-2) and diploid reference (2x-1) by methylation-sensitive amplification polymorphism (MSAP). The total methylation level of q2x-1, 4x-1, and 4x-2 was higher than that of 2x-1. The total methylation level of q2x-1 was higher than that of 4x-1 and 4x-2. Compared with 2x-1, the methylation pattern was mainly maintenance of methylation in q2x-1, 4x-1, and 4x-2, but the trend of demethylation of q2x-1, 4x-1, and 4x-2 was greater than that of methylation. Compared with q2x-1, the methylation pattern in 4x-1 and 4x-2 is also mainly maintaining methylation, but the trend of demethylation in 4x-1 and 4x-2 is less than that of methylation. The results of BLAST homology comparison and expression verification showed that the ploidy change of plants may affect the expression of genes related to adenosine triphosphate binding, RNA binding, arginine biosynthesis, and cell shape regulation, as well as promoting the polarization growth of leaf epidermis, auxin mediation, and RNA modification by causing a change of DNA methylation. This study provides a theoretical basis for studying the molecular mechanisms of plant ploidy change from the perspective of epigenetics. [ABSTRACT FROM AUTHOR]